Recombinant microorganisms and uses therefor

Patent metadata

Field	Value
Publication number	US-9410130-B2
Application number	US-201213403972-A
Country	US
Kind code	B2
Filing date	Feb 23, 2012
Priority date	Feb 25, 2011
Publication date	Aug 9, 2016
Grant date	Aug 9, 2016

Abstract

The invention provides, inter alia, methods for the production of acetone, isopropanol and/or precursors of acetone and/or isopropanol by microbial fermentation of substrates comprising CO, genetically modified microorganisms of use in such methods, nucleic acids suitable for preparation of genetically modified microorganisms, a novel alcohol dehydrogenase and nucleic acids encoding same.

First claim

The invention claimed is:

1. A recombinant microorganism comprising an exogenous thiolase (EC 2.3.1.9), an exogenous CoA transferase (EC 2.8.3.9), and an exogenous decarboxylase (EC 4.1.1.4 or EC 4.1.1.74), wherein the recombinant microorganism is derived from a parental microorganism selected from the group consisting of Clostridium autoethanogenum, Clostridium ljungdahlii , and Clostridium ragsdalei , wherein the recombinant microorganism produces isopropanol.

2. The recombinant microorganism of claim 1 , wherein the thiolase is Clostridium acetobutylicum ThlA or Clostridium beijerinckii ThlA.

3. The recombinant microorganism of claim 2 , wherein the Clostridium acetobutylicum ThlA comprises SEQ ID NO: 42.

4. The recombinant microorganism of claim 1 , wherein the CoA transferase is Clostridium acetobutylicum CtfA and CtfB or Clostridium beijerinckii CtfA and CtfB.

5. The recombinant microorganism of claim 4 , wherein the Clostridium beijerinckii CtfA comprises SEQ ID NO: 43 and CtfB comprises SEQ ID NO: 44.

6. The recombinant microorganism of claim 1 , wherein the decarboxylase is acetoacetate decarboxylase (EC 4.1.1.4).

7. The recombinant microorganism of claim 6 , wherein the acetoacetate decarboxylase is Clostridium acetobutylicum Adc or Clostridium beijerinckii Adc.

8. The recombinant microorganism of claim 7 , wherein the Clostridium beijerinckii Adc comprises SEQ ID NO: 45.

9. The recombinant microorganism of claim 1 , wherein the decarboxylase is ketoisovalerate decarboxylase (EC 4.1.1.74).

10. The recombinant microorganism of claim 9 , wherein the ketoisovalerate decarboxylase is Lactococcus lactis KivD.

11. The recombinant microorganism of claim 10 , wherein the Lactococcus lactis KivD comprises SEQ ID NO: 73.

12. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises an endogenous or exogenous alcohol dehydrogenase.

13. The recombinant microorganism of claim 12 , wherein the endogenous or exogenous alcohol dehydrogenase is alcohol dehydrogenase (Adh) (EC 1.1.1.2) or alcohol dehydrogenase (Adh2) (EC 1.1.1.1).

14. The recombinant microorganism of claim 12 , wherein the exogenous alcohol dehydrogenase is Saccharomyces cerevisiae Adh2 (EC 1.1.1.1).

15. The recombinant microorganism of claim 1 , wherein the recombinant microorganism is a carboxydotrophic and acetogenic bacterium.

16. The recombinant microorganism of claim 1 , wherein the parental microorganism does not produce acetone or isopropanol.

17. The recombinant microorganism of claim 1 , wherein the parental microorganism is Clostridium autoethanogenum or Clostridium ljungdahlii.

18. The recombinant microorganism of claim 1 , wherein the parental microorganism is Clostridium autoethanogenum DSM23693 or Clostridium ljungdahlii DSM13528.

19. The recombinant microorganism of claim 1 , wherein the recombinant microorganism consumes a gaseous substrate comprising one or more of CO, CO 2 , and H 2 .

20. The recombinant microorganism of claim 19 , wherein the gaseous substrate comprises 20-70% CO.

21. The recombinant microorganism of claim 19 , wherein the gaseous substrate is derived from an industrial process selected from the group consisting of ferrous metal products manufacturing, non-ferrous products manufacturing, petroleum refining, coal gasification, biomass gasification, electric power production, carbon black production, and coke manufacturing.

Assignees

• Koepke Michael
• Simpson Sean
• Liew Fungmin
• Chen Wendy
• Lanzatech New Zealand Ltd

Inventors

• Koepke Michael
• Simpson Sean
• Liew Fungmin
• Chen Wendy

Classifications

• C12Y401/01004

  Acetoacetate decarboxylase (4.1.1.4) · CPC title

• C12Y208/03009

  Butyrate--acetoacetate CoA-transferase (2.8.3.9) · CPC title

• C12N9/1029

  transferring groups other than amino-acyl groups (2.3.1) · CPC title

• C12N9/13

  transferring sulfur containing groups (2.8) · CPC title

• C12Y203/01009

  Acetyl-CoA C-acetyltransferase (2.3.1.9) · CPC title